Folded globular proteins are attractive building blocks for biomaterials as their robust structures carry out diverse biological functions. These biomaterials are ideal to study the translation of molecular properties to multi-molecular assemblies.
Hierarchical assemblies
of proteins exhibit a wide-range of material
properties that are exploited both in nature and by artificially by
humankind. However, little is understood about the importance of protein
unfolding on the network assembly, severely limiting opportunities
to utilize this nanoscale transition in the development of biomimetic
and bioinspired materials. Here we control the force lability of a
single protein building block, bovine serum albumin (BSA), and demonstrate
that protein unfolding plays a critical role in defining the architecture
and mechanics of a photochemically cross-linked native protein network.
The internal nanoscale structure of BSA contains “molecular
reinforcement” in the form of 17 covalent disulphide “nanostaples”,
preventing force-induced unfolding. Upon addition of reducing agents,
these nanostaples are broken rendering the protein force labile. Employing
a combination of circular dichroism (CD) spectroscopy, small-angle
scattering (SAS), rheology, and modeling, we show that stapled protein
forms reasonably homogeneous networks of cross-linked fractal-like
clusters connected by an intercluster region of folded protein. Conversely,
in situ
protein unfolding results in more heterogeneous
networks of denser fractal-like clusters connected by an intercluster
region populated by unfolded protein. In addition, gelation-induced
protein unfolding and cross-linking in the intercluster region changes
the hydrogel mechanics, as measured by a 3-fold enhancement of the
storage modulus, an increase in both the loss ratio and energy dissipation,
and markedly different relaxation behavior. By controlling the protein’s
ability to unfold through nanoscale (un)stapling, we demonstrate the
importance of
in situ
unfolding in defining both
network architecture and mechanics, providing insight into fundamental
hierarchical mechanics and a route to tune biomaterials for future
applications.
The structure of aggregates and gels formed by heat-denatured whey protein isolate (WPI) has been studied at pH 7 and different ionic strengths using light scattering and turbidimetry. The results were compared with those obtained for pure beta-lactoglobulin (beta-Lg). WPI aggregates were found to have the same self-similar structure as pure beta-Lg aggregates. WPI formed gels above a critical concentration that varied from close to 100 g/L in the absence of added salt to about 10 g/L at 0.2 M NaCl. At low ionic strength (<0.05 M NaCl) homogeneous transparent gels were formed, while at higher ionic strength the gels became turbid but had the same self-similar structure as reported earlier for pure beta-Lg. The length scale characterizing the heterogeneity of the gels increased exponentially with increasing NaCl concentration for both WPI and pure beta-Lg, but the increase was steeper for the former.
We describe the possibility to create solid-like protein samples whose structural and mechanical properties can be varied and tailored over an extremely large range in a very controlled way through an arrested spinodal decomposition process. We use aqueous lysozyme solutions as a model globular protein system. A combination of video microscopy, small-angle neutron and X-ray scattering and reverse Monte Carlo modeling is used to characterize the structure of the bicontinuous network with two coexisting phases of a dilute protein solution and a glassy or arrested dense protein backbone at all relevant length scales. Rheological measurements are then used to determine the complex mechanical response of these protein gels as a function of protein concentration and quench temperature. While in particular the origin of the dependence of the mechanical properties on quench depth and concentration is not well understood currently, it seems ultimately connected to the particular bicontinuous structure of the arrested spinodal network created by the interplay between the early stage of a spinodal decomposition and the position of the glass line. We then generalize this behavior and discuss how this could open up new routes to prepare gel-like food systems with adjustable structural and mechanical properties. We present results from a first feasibility study where we use a depletion interaction caused by the addition of small non-adsorbing polymers to suspensions of casein micelles in order to create food gels with tunable structural and mechanical properties through an arrested spinodal decomposition process.
Investigating proteins with techniques such as NMR or neutron scattering frequently requires the partial or complete substitution of D2O for H2O as a solvent, often tacitly assuming that such a solvent substitution does not significantly alter the properties of the protein. Here, we report a systematic investigation of the solvent isotope effect on the phase diagram of the lens protein γB-crystallin in aqueous solution as a model system exhibiting liquid-liquid phase separation. We demonstrate that the observed strong variation of the critical temperature Tc can be described by the extended law of corresponding states for all H2O/D2O ratios, where scaling of the temperature by Tc or the reduced second virial coefficient accurately reproduces the binodal, spinodal, and osmotic compressibility. These findings highlight the impact of H2O/D2O substitution on γB-crystallin properties and warrant further investigations into the universality of this phenomenon and its underlying mechanisms.
We investigate the charge transport physics of a previously unidentified class of electron-deficient conjugated polymers that do not contain any single bonds linking monomer units along the backbone but only double-bond linkages. Such polymers would be expected to behave as rigid rods, but little is known about their actual chain conformations and electronic structure. Here, we present a detailed study of the structural and charge transport properties of a family of four such polymers. By adopting a copolymer design, we achieve high electron mobilities up to 0.5 cm2 V−1 s−1. Field-induced electron spin resonance measurements of charge dynamics provide evidence for relatively slow hopping over, however, long distances. Our work provides important insights into the factors that limit charge transport in this unique class of polymers and allows us to identify molecular design strategies for achieving even higher levels of performance.
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